氧化应激相关的LncRNA和miRNA相互作用在糖尿病视网膜病变发病中的作用

Oxidative stress is one of the pathogenic mechanism of diabetic retinopathy (DR). Long non coding RNA (LncRNA) is a class of RNA with variety of biological function. Recent studies have shown that many lncRNAs were aberrantly expressed in the retinas of diabetic mice model. However, the function and mechanism of the aberrantly expressed lncRNA remains unclear. We previously found that oxidative stress altered multiple lncRNA expression in human umbilical vein endothelial cells (HUVECs) by the mixed sample sequencing. This project will perform lncRNAs expression profiling of oxidative stress-induced HUVECs using microarray analysis to identify the oxidative stress-related lncRNAs. The effect of candidate lncRNAs on the endothelial function will be investigated in the oxidative stress HUVECs and the retinas of diabetic mice model by overexpression and knockdown of the lncRNAs.Further more, based on the bioinformatics analysis, the lncRNA which is probably with the combination of miRNA (screened by other project) will be selected, and the interaction between lncRNAs and microRNA will be investigated by immunoprecipitation analysis and the report gene analysis.At last, effects of the interaction between lncRNAs and microRNA on the function of HUVECs will be identified by angiogenesis assay permeability assay. This project could explore the role and molecular mechanism of lncRNA in the pathogenesis of DR from the aspect of oxidative stress, and provide a new idea for the prevention and treatment of DR.

氧化应激是糖尿病视网膜病变（DR）的重要发病机制之一。长链非编码RNA（LncRNA）具有广泛的生物学功能。研究证实，DR动物视网膜中LncRNA表达明显改变，但其功能和分子机制尚未阐明。我们前期通过对H2O2刺激前后人脐静脉内皮细胞(HUVECs)RNA混样测序，发现氧化应激可导致多种LncRNA表达改变。本项目拟建立氧化应激细胞模型，采用lncRNA芯片分析，筛选与氧化应激密切相关的lncRNAs；在氧化应激细胞模型和DR动物模型上通过敲低和过表达lncRNA，观察候选lncRNA对内皮功能的影响；结合生物信息学分析，筛选出可能与目标lncRNA相互作用的miRNA，利用免疫沉淀、报告基因分析等技术，揭示lncRNA和miRNA的相互作用，进一步明确二者的相互作用对HUVECs功能的影响。本研究从氧化应激角度探讨LncRNA 在DR发病中的作用及分子机制，为DR的防治提供新思路。